title: [Development of selective detection method for 5-formyl-2'-deoxyuridine in DNA using a fluorogenic reagent].
abstract: It is important that various lesions in DNA were detected selectively and conveniently to know mechanisms of carcinogenicity and/or aging of cells. However, most detection methods of DNA lesion are complicated and take a long time for enzymatic hydrolysis and analysis by HPLC and/or mass spectrometry. This review shows the new concept for detection of DNA lesion by "fluorogenic reagent". Inspired by the unique bis-heteroaryl structure of luciferin and 5-heteroaryl-2'-deoxyuridine having good fluorescence properties, we designed and synthesized fluorogenic reagent 4,5-dimethoxy-2-aminothiophenol for a selective and convenient detection for 5-formyl-2'-deoxyuridine, which is generated in yields comparable to that of 2'-deoxy-8-oxoguanosine, in DNA. Generated 5-(5,6-dimethoxybenzothiazol-2-yl)-2'-deoxyuridine has a high quantum yield and larger Stokes shift in aqueous solution. This derivatization of 5-formyl-2'-deoxyuridine in oligodeoxynucleotide occurred quickly and quantitatively. The fluorogenic reagent was also revealed to detect 5-formyl-2'-deoxyuridine in γ-irradiated calf thymus DNA with irradiation dose dependent manner. Thus, our fluorogenic strategy enables to selective and convenient detection of lesion in DNA exposed to various forms of oxidative stress.
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokkaido University.
authors.forename: Kousuke
authors.initials: K
authors.lastname: Sato
mesh_terms: D000814:Aniline Compounds / Q000138:chemical synthesis*; D000818:Animals; D004247:DNA / Q000737:chemistry*; D004249:DNA Damage; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000032:analysis; D015195:Drug Design; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry; D005470:Fluorometry / Q000379:methods*; D018384:Oxidative Stress / Q000235:genetics*
chemical_list: C588011:4,5-dimethoxy-2-aminothiophenol; C402104:5-formyl-2'-deoxycytidine; D000814:Aniline Compounds; D005456:Fluorescent Dyes; D003841:Deoxycytidine; D004247:DNA